Hypericum Triquetrifolium Callus Cultures a Potential Source of Phenolics and Flavonoids


1 Hoshyar A. Azeez, 2 Kadhim M. Ibrahim
1 Biology Dept., Faculty of Science and Science Education, School of Science, Sulaimani University, 2 Biotechnology Dept., College of Sciences, Al-Nahrain University, Baghdad,






Abstract:
An experiment was conducted to induce and increase the production of some
secondary metabolites in tissue cultures of Hypericum triquetrifolium Turra using some
biotechnological approaches. Callus was initiated on leaf discs cultured on Murashiege
and Skoog medium (MS) supplemented with Thidiazuron (TDZ) at the concentrations
1.0, 1.25, 1.5, 2.0, or 2.5 mg L-1 and indole-3-acetic acid (IAA) at 0.5 mgL-1; callus was
initiated on stem and root explants on MS medium supplemented with 1.25 mgL-1 6-
benzyl-aminopurine (BAP) and 0.5 mg L-1of indol acetic acid (IAA). Results showed
that the combination of TDZ at 2 mg L-1 with 0.5 mg L-1 IAA was the most effective in
inducing callus formation on leaf explants. The same combination was used for callus
maintenance. HPLC was used to determine the type and quantity of secondary
metabolites in comparison with standards. The differences in phytochemical
composition among leaf (L), stem (S), and root (R) were investigated. The highest levels
of phenolic compounds and flavonoids were present in L compared to S and R extracts,
except the production of rutin and hypersoid in S which was more than L and R. R
accumulated larger amounts of chlorgenic and catechin than L and S, while the caffeic
acid and tannic acid production in cell suspension cultures derived from leaf (LCs)
increased significantly up to 5.6 and 4.2 folds respectively compared to L and callus
derived from leaf (LC). P-OH-benzoic acid, caffeic acid, quercitin and tannic acid were
absent in S, whereas all of these compounds were present in callus derived from stem
(SC) and cell suspension cultures derived from stem (SCs), but in roots only p-OH-
benzoic acid was not produced.

Key words: Hypericum triquetrifolium Turra, Callus, suspension cultures, Phenolics,
Flavonoids.



References

(1) Al-Mukhtar, J.A.H. (1975). Hypericum plant. Directorate plant. Bulletin No. 231.
Ministry of Agriculture and Agrarian Reform, Iraq. pp. 2-15.
(2) Barnes, J., Anderson, L.A. and Phillipson, J.D. (2001). St. John's Wort (Hypericum
perforatum L.) A review of its chemistry, pharmacology and clinical properites. J. Pharm.
Pharmcol., 53:583-600.
(3) Cirak, C., Bertoli, A., Pistelli, L. and Seyis, F. (2010). Essential oil composition and
variability of Hypericum perforatum from wild populations of northern Turkey. Pharm. Biol.,
48:906-914.
(4) Hosni, K., Msaada, K., Taarit, M.B. and Marzouk, B. (2011). Phonological variations of
secondary metabolites from Hypericum triquetrifolium Turra. Biochemical Systematics and
Ecology, 39:43-50.
(5) Vijaya Sree, N., Udayasri, P.V.V., Aswani, K.Y., Ravi, B.B., Phani, K.Y. and Vijay, V.M.
(2010). Advancements in the production of secondary metabolites. J. of Natural Products,
3:112-123.
(6) Oluk, E.A., Orhan, S., Karakas, O., Cakir, A. and Gonuz, A. (2010). High efficiency
indirect shoot regeneration and hypericin content in ebryogenic callus of Hypericum
triqutrifolium Turra. Afric. J. of Biotechnology, 9(15): 2229-2233.
(7) Cirak, C. (2007). Seed germination protocols for Ex situ conservation of some Hypericum
species from Turkey. Am. J. Plant Physiol., 2:287-294.
(8) Oluk, E.A. and Orhan, S. (2009). Thidiazuron induced micropropagation of Hypericum
triqutrifolium Turra. Afric. J. of Biotechnology, 8(15): 3506-3510.
(9) Gopi, G. and Vatsala, T.M. (2006). In vitro studies on effects of plant growth regulators
on callus and cell suspension culture biomass yield from Gymnemasylvestre R.Br. African J.
of Biotech., 5 (12), 1215-1219.
(10) Tasis, E.C., Boeren, S., Exarchou, V., Troganis, A.N., Vervoort, J.and Gerothanassis,
I.P. (2007). Identification of the major constituents of Hypericum perforatum by
LC\SPE\NMR and \or LC\MS. Phytochemistry, 68: 383-393.
(11) Duncan, D.B. (1995). Multiple range and multiple F-test, Biometrics., 11:1-42.
(12) Rojbayani, S.A.K. (2007). Induction and stimulation of some phytochemicals from
Hypericum perforatum L. in vitro. PhD Dissertation. Al- Mustansiriya University. pp. 59-67.
(13) Khan, T., Krupadanam, D. and Anwar, S.Y. (2008). The role of phytohormone on the
production of berberine in the calli cultures of an endangered medicinal plant, turmeric
(Coscinium fenestratum l.) AfricanJ. of Biotech., 7 (18):244-3246.